• Development and Reproduction
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• v.16 no.1
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• pp.31-38
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• 2012

Kisspeptin has been implicated in the process of puberty onset in various animal groups. This peptide is encoded by a gene, Kiss1 in avian and mammalian species. Contrary to these higher vertebrates, however, fish appeared to have another gene, Kiss2 that also codes for the precursor peptide of kisspeptin. To figure out biological significance of this gene during the puberty onset in fish, the expression profile of Kiss2 gene was investigated in the brain of Nile tilapia together with genes of GPR54, GnRH receptorI (rGnRHI) and GTH subunits ($LH{\beta}$ and $FSH{\beta}$). Expression of Kiss2 mRNA significantly increased at 2 weeks post hatch (wph) and 13 wph ($P$<0.05). This increase coincided with the increases of GPR54 and rGnRH I gene expression. Detection of $LH{\beta}$ and $FSH{\beta}$ subunit gene expression was possible later than 13 wph, indicating the activation of gonadotrophs in the pituitary. Data obtained from this study strongly suggest that, in addition to Kiss1 gene, Kiss2 gene is deeply associated with the onset of puberty by the activation of hypothalamus pituitary gonadal axis in Nile tilapia.

• Development and Reproduction
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• v.28 no.1
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• pp.21-28
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• 2024

Present study aimed to investigate the temporal changes in expression of some reproductive hormones in testis, originally found in hypothalamus and pituitary. Rats were sacrificed on postnatal day 23 (PND23; immature), pubertal (PND53) and PND 81 (young adult). The testicular RNAs were extracted, and semi-quantitative PCRs for gonadotropin-releasing hormone (GnRH), kisspeptin 1 (KiSS1), pituitary adenylate cyclase-activating polypeptide (PACAP), LH subunits and LH receptor were performed. Transcript levels of GnRH and KiSS1 at PND23 were significantly higher than levels of PND53 and PND81 (p<0.001). PACAP mRNA level at PND23 was significantly lower than those of PND53 and PND81 (p<0.001). The mRNA levels of both testis type and pituitary type luteinizing hormone β subunit (tLHβ and pLHβ, respectively) at PND23 were significantly lower than levels of PND53 and PND81 (p<0.001). The mRNA level of glycoprotein hormone common alpha subunit (Cgα) at PND23 was significantly lower than those of PND53 and PND81 (p<0.001). Present study revealed the intratesticular expression of KiSS1 and GnRH showed a very similar trend while the expression of PACAP in the testis showed reversed pattern. The expressions of LHβ subunits (tLHβ and pLHβ) were very low during immature stage then increased significantly during puberty and early adulthood. Our attempt to study the local role(s) of intratesticular factors will be helpful to achieve precise understanding on the testis physiology and pathology.

• Journal of Life Science
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• v.11 no.2
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• pp.103-107
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• 2001

The equine chorionic gonadotropin (eCG) subunits $\alpha$ and ${\beta}$ are transcribed from different genes and associate noncovalently to form the bioactive eCG heterodimer. Dimerization is rate limiting for eCG secretion, and dissociation leads to hormone inactivation. The correct conformation of the heterodimer is alto important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduction. To determine whether ${\alpha}$ and ${\beta}$ subunits can be synthesized as a single polypeptide chain (tethered-eCG) and also display biological activity, the tethered-eCG molecule by fusing the carboxyl terminus of the eCG ${\beta}$-subunit to the amino terminus of the af-subunit was construe-ted and transfected into chinese hamster ovary (CHO-Kl) cells. LH- and FSH-like activities were assayed in terms of testosterone production and aromatase activity in primary cultured rat Leydig cells and granulosa cells, respectively. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG ${\alpha}$/${\beta}$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG ${\alpha}$/${\beta}$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-K1 cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models of FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.4
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• pp.106-118
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• 2005

Purpose : Deoyeongjeon(DYJ : 大營煎) is used in female infertility, caused by　ovulation disorder. so this study is to examine what are the effects of the Deoyeongjeon(DYJ) on the vulation and Ovary in Rats Methods : 4weeks Female Sprague-Dawley 12 rats of weighting 160-l80g, were divided into three groups including the DYJ oral administration(4ml/kg) groups(4heads) and DYJ oral administration(8m/lkg) groups(4heads). then we observed changes in the serum concentrations of FSH, LH, and estradiol(E2) and the histological changes of ovary and the immunohistochemical staining for progesterone receptor in ovary of rats. Results : 1. Blood FSH level significantly increased in experimental group as compared with control group. 2. In blood LH level, experimental group showed no efficacy as compared with control group. 3. In blood estradiol(E2) level, experimental group showed no efficacy as compared with control group. 4. In histological observations of ovary, ovulation increased in experimental group as compared with control group, which showed no efficacy. 5. In observations of immunohistochemical staining for progesterone receptor in ovary, immunohistochemical staining score (ISS) of atretic follicles significantly showed a tendency to decrease in experimental group as compared with control group. Conclusion : DYJ influences the pituitary gland and ovary to increase the ovulation of rats.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.54-54
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• 2001

Human chorionic gonadotropin (hCG) is a member of the glycoprotein hormone family which includes FSH, hCG, TSH. These hormone family is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a hormone specific $\beta$-subunit. The correct conformation of the heterodimer is also important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduciton. To determine $\alpha$ and $\beta$-subunits can be synthesized as a single polypeptide chain (tethered-hCG) and also display biological activity, the tethered-hCG molecule by fusing the carboxyl terminus of the hCG $\beta$-subunit to the amino terminus of the $\alpha$-subunit was constructed and transfected into chinese hamster ovary (CHO-K1) cells. We also constructed C-terminal deletion mutants (D9l, D89, D88, D87, D86, D84, D83) of single chain hCG to determine the biological function (secretion, LH-activity, receptor binding, cAMP production) of these mutants. Between six and eight stably transfected pools of cells expressing wild type and mutant hCGs were selected for neomycin resistant. The hCGs secreted by the stably transfected cells into serum-free media were collected and quantified by radioimmunoassay, as described in protocol (DPC(hCG IRMA). LH activity was in terms of testosterone production and aromatase activity in primary cultured rat Leydig cells. The tethered-wthCG was efficiently secreted and showed similar LH-like activity to the dimeric hCG. The D83hCG mutant was not detected in this assay. It is suggest that hCG C-terminal part is very important for hCG secretion. Now, we checking the LH-like activity of these mutant hCGs. These data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.207-212
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• 2020

Objective: Glutamate ionotropic receptor AMPA type subunit 1 (GRIA1) is a subunit of a ligand-gated ion channel that regulates the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by controlling the release of gonadotropin-releasing hormone. Few studies have investigated the association between the GRIA1 gene and human infertility. This study evaluated the association of the GRIA1 rs548294 C > T and rs2195450 G > A polymorphisms with the ovarian response to human menopausal gonadotropin (HMG) in Iranian women. Methods: One hundred women with histories of at least 1 year of infertility were included. On the second day of menstruation, patients were injected with HMG; on the third day, blood samples were collected. After hormonal analysis, the GRIA1 rs548294 C > T and rs2195450 G > A genotypes of samples were identified via the restriction fragment length polymorphism method, and on day 9, the number of follicles was assessed via ultrasound. Results: For the GRIA1 rs548294 C > T and rs2195450 G > A single nucleotide polymorphisms, the subjects with CT and GG genotypes, respectively, displayed the highest mean FSH level, LH level, and number of follicles on day 9 of the menstrual cycle (p< 0.05). Significant positive correlations were observed between LH and FSH (p< 0.01), LH and follicle count (p< 0.01), FSH and age (p< 0.05), follicle count and age (p= 0.048), and FSH and follicle count (p< 0.01). Conclusion: This study showed a significant relationship between GRIA1 polymorphisms and ovarian response to the induction of ovulation. Therefore, determining patients' GRIA1 genotype may be useful for improving treatment and prescribing suitable doses of ovulation-stimulating drugs.

• Clinical and Experimental Reproductive Medicine
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• v.11 no.2
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• pp.27-39
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• 1984

The purpose of this experiment was to investigate the effects of experimental cryptorchidism on the levels of testicular FSH, LH receptor. One hundred and thirty two male rats weighing 100${\pm}$10g, 45 days old, were divided into three groups of control, bilateral cryptorchidism and unilateral cryptorchidism which was subdivided into two groups of scrotal testis and abdominal testis. Thirty to forty two heads of rats were allotted to each group. Five to eight rats from each group were randomely sacrificed at 1, 2, 3, 4 and 5 weeks after surgical induction of cryptorchidism. Testis was removed for assay of FSH, LH receptors. The results obtained were as follows. 1. The levels of FSH receptors per 10mg testicular tissue in abdominal testis of unilateral cryptorchid group and bilateral cryptorchid group tended to decrease gradually toward 5th week and these levels at 2nd, 3rd, 4th and 5th week were significantly lower than those levels of control group and scrotal testis of unilateral cryptorchid group. On the other hand; the ranges of affinities of FSH receptors of abdominal testis in unilateral and bilateral cryptorchid group were 2.2 - 6.67 ${\times}10^{11}M^{-1}$ and 3.6 - 4.8 ${\times}10^{11}M^{-1}$ respectively, and these values were slightly higher than those of control group (1.43 - 3.98 ${\times}10^{11}M^{-1}$) or those of scrotal testis in unilateral cryptorchid group (1.41 - 3.25 ${\times}10^{11}M^{-1}$). Especially at I and 4 weeks after treatment the affinities of abdominal testis in unilateral and bilateral cryptorchid group were significantly higher than those of other group (P <0.01). 2. The levels of LH receptors in 10mg testicular tissue tended to increase with elapse of time in control and in scrotal testis of unilateral cryptorchid group. But the levels of LH receptors of abdominal testis in unilateral and bilateral cryptorchid group tended to decrease after treatment and were significantly lower than control group at 1 (P <0.01),2,4 and 5 weeks after treatment (P < 0.05). The affinities of LH receptors were not significantly different among treatments except those of bilateral cryptorchid group and abdominal testis of unilateral cryptorchid group being significantly higher than that of the control at 4 weeks after treatment.

• Development and Reproduction
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• v.25 no.4
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• pp.225-234
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• 2021

The present study aimed to investigate the mechanism of cell surface receptor loss by two constitutively activating mutants (designated L469R, and D590Y) and two inactivating mutants (D417N and Y558F) of the luteinizing hormone receptor (LHR) in the Japanese eel Anguilla japonica, known to naturally occur in human LHR transmembrane domains. We investigated cell surface receptor loss using an enzyme-linked immunosorbent assay in HEK 293 cells. The expression level of wild-type eel LHR was considered to be 100%, and the expression levels of L469R and D417N were 97% and 101%, respectively, whereas the expression levels of D590Y and Y558F slightly increased to approximately 110% and 106%, respectively. The constitutively activating mutants L469R and D590Y exhibited a decrease in cell surface loss in a manner similar to that of wild-type eel LHR. The rates of loss of cell surface agonist-receptor complexes were observed to be very rapid (2.6-6.2 min) in both the wild-type eel LHR and activating mutants. However, cell surface receptor loss in the cells expressing inactivating mutants D417N and Y558F was slightly observed in the cells expressing inactivating mutants D417N and Y558F, despite treatment with a high concentration of agonist. These results provide important information on LHR function in fish and the regulation of mutations of highly conserved amino acids in glycoprotein hormone receptors.

• 대한생식의학회:학술대회논문집
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• 1999.11a
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• pp.105-106
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• 1999

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.83-83
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• 2003